PDLIM5 CRISPR Activation Plasmid (h): sc-405340-ACT

PDLIM5 encodes a PDZ and LIM domain–containing scaffold protein that localizes to actin-associated structures and links cytoskeletal organization to signal transduction. In human cells, PDLIM5 participates in regulation of actin dynamics, cell adhesion, and mechanosensitive pathways by coordinating protein–protein interactions at stress fibers and Z-disc–like assemblies. Through these functions, it influences processes such as cell morphology, migration, and contractility, and interfaces with kinase signaling networks that respond to cytoskeletal cues. Altered PDLIM5 expression and pathway activity have been studied in the context of cardiovascular and neuropsychiatric disease biology, supporting its utility as a node for investigating cytoskeleton-coupled signaling mechanisms.

PDLIM5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PDLIM5 expression without altering the underlying DNA sequence.

PDLIM5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PDLIM5 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the PDLIM5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PDLIM5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PDLIM5 locus and enabling the study of PDLIM5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PDLIM5 pathway restoration in tumor cells with silenced or reduced PDLIM5 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.